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Yorodumi- PDB-6gsn: Structure of a partial yeast 48S preinitiation complex in closed ... -
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-Basic information
Entry | Database: PDB / ID: 6gsn | |||||||||
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Title | Structure of a partial yeast 48S preinitiation complex in closed conformation | |||||||||
Components |
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Keywords | RIBOSOME / translation / initiation factors / 40S / eIF1A / eIF3 / eIF2 / tRNAi / 48S PIC / small ribosome subunit | |||||||||
Function / homology | Function and homology information formation of translation initiation ternary complex / Recycling of eIF2:GDP / Cellular response to mitochondrial stress / eukaryotic translation initiation factor 3 complex, eIF3e / ABC-family proteins mediated transport / eukaryotic translation initiation factor 3 complex, eIF3m / methionyl-initiator methionine tRNA binding / incipient cellular bud site / translation reinitiation / eukaryotic translation initiation factor 2 complex ...formation of translation initiation ternary complex / Recycling of eIF2:GDP / Cellular response to mitochondrial stress / eukaryotic translation initiation factor 3 complex, eIF3e / ABC-family proteins mediated transport / eukaryotic translation initiation factor 3 complex, eIF3m / methionyl-initiator methionine tRNA binding / incipient cellular bud site / translation reinitiation / eukaryotic translation initiation factor 2 complex / eukaryotic translation initiation factor 3 complex / formation of cytoplasmic translation initiation complex / multi-eIF complex / cytoplasmic translational initiation / protein-synthesizing GTPase / eukaryotic 43S preinitiation complex / formation of translation preinitiation complex / eukaryotic 48S preinitiation complex / positive regulation of translational fidelity / Formation of the ternary complex, and subsequently, the 43S complex / Translation initiation complex formation / Ribosomal scanning and start codon recognition / Formation of a pool of free 40S subunits / L13a-mediated translational silencing of Ceruloplasmin expression / ribosomal small subunit binding / 90S preribosome / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of translational fidelity / translation regulator activity / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / translation initiation factor binding / translation initiation factor activity / rescue of stalled ribosome / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / translational initiation / protein kinase C binding / maintenance of translational fidelity / cytoplasmic stress granule / modification-dependent protein catabolic process / protein tag activity / rRNA processing / ribosomal small subunit biogenesis / double-stranded RNA binding / small ribosomal subunit rRNA binding / ribosome binding / ribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / cytoplasmic translation / cytosolic large ribosomal subunit / rRNA binding / ribosome / protein ubiquitination / structural constituent of ribosome / translation / ribonucleoprotein complex / positive regulation of protein phosphorylation / GTPase activity / mRNA binding / ubiquitin protein ligase binding / nucleolus / GTP binding / protein kinase binding / RNA binding / zinc ion binding / identical protein binding / nucleus / metal ion binding / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) Kluyveromyces lactis (yeast) | |||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 5.75 Å | |||||||||
Authors | Llacer, J.L. / Hussain, T. / Ramakrishnan, V. | |||||||||
Funding support | United Kingdom, 2items
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Citation | Journal: Nucleic Acids Res / Year: 2021 Title: Large-scale movement of eIF3 domains during translation initiation modulate start codon selection. Authors: Jose L Llácer / Tanweer Hussain / Jinsheng Dong / Laura Villamayor / Yuliya Gordiyenko / Alan G Hinnebusch / Abstract: The eukaryotic initiation factor 3 (eIF3) complex is involved in every step of translation initiation, but there is limited understanding of its molecular functions. Here, we present a single ...The eukaryotic initiation factor 3 (eIF3) complex is involved in every step of translation initiation, but there is limited understanding of its molecular functions. Here, we present a single particle electron cryomicroscopy (cryo-EM) reconstruction of yeast 48S ribosomal preinitiation complex (PIC) in an open conformation conducive to scanning, with core subunit eIF3b bound on the 40S interface near the decoding center in contact with the ternary complex eIF2·GTP·initiator tRNA. eIF3b is relocated together with eIF3i from their solvent interface locations observed in other PIC structures, with eIF3i lacking 40S contacts. Re-processing of micrographs of our previous 48S PIC in a closed state also suggests relocation of the entire eIF3b-3i-3g-3a-Cter module during the course of initiation. Genetic analysis indicates that high fidelity initiation depends on eIF3b interactions at the 40S subunit interface that promote the closed PIC conformation, or facilitate the relocation of eIF3b/eIF3i to the solvent interface, on start codon selection. #1: Journal: Mol Cell / Year: 2015 Title: Conformational Differences between Open and Closed States of the Eukaryotic Translation Initiation Complex. Authors: Jose L Llácer / Tanweer Hussain / Laura Marler / Colin Echeverría Aitken / Anil Thakur / Jon R Lorsch / Alan G Hinnebusch / V Ramakrishnan / Abstract: Translation initiation in eukaryotes begins with the formation of a pre-initiation complex (PIC) containing the 40S ribosomal subunit, eIF1, eIF1A, eIF3, ternary complex (eIF2-GTP-Met-tRNAi), and ...Translation initiation in eukaryotes begins with the formation of a pre-initiation complex (PIC) containing the 40S ribosomal subunit, eIF1, eIF1A, eIF3, ternary complex (eIF2-GTP-Met-tRNAi), and eIF5. The PIC, in an open conformation, attaches to the 5' end of the mRNA and scans to locate the start codon, whereupon it closes to arrest scanning. We present single particle cryo-electron microscopy (cryo-EM) reconstructions of 48S PICs from yeast in these open and closed states, at 6.0 Å and 4.9 Å, respectively. These reconstructions show eIF2β as well as a configuration of eIF3 that appears to encircle the 40S, occupying part of the subunit interface. Comparison of the complexes reveals a large conformational change in the 40S head from an open mRNA latch conformation to a closed one that constricts the mRNA entry channel and narrows the P site to enclose tRNAi, thus elucidating key events in start codon recognition. | |||||||||
History |
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-Structure visualization
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Structure viewer | Molecule: MolmilJmol/JSmol |
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PDBx/mmCIF format | 6gsn.cif.gz | 2.2 MB | Display | PDBx/mmCIF format |
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PDB format | pdb6gsn.ent.gz | Display | PDB format | |
PDBx/mmJSON format | 6gsn.json.gz | Tree view | PDBx/mmJSON format | |
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-Validation report
Summary document | 6gsn_validation.pdf.gz | 1.8 MB | Display | wwPDB validaton report |
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Full document | 6gsn_full_validation.pdf.gz | 1.9 MB | Display | |
Data in XML | 6gsn_validation.xml.gz | 208.9 KB | Display | |
Data in CIF | 6gsn_validation.cif.gz | 355.9 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/gs/6gsn ftp://data.pdbj.org/pub/pdb/validation_reports/gs/6gsn | HTTPS FTP |
-Related structure data
Related structure data | 0058MC 0057C 6gsmC C: citing same article (ref.) M: map data used to model this data |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
-RNA chain , 3 types, 3 molecules 231
#1: RNA chain | Mass: 579239.625 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: GenBank: 49642208 |
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#20: RNA chain | Mass: 4321.563 Da / Num. of mol.: 1 / Source method: obtained synthetically Source: (synth.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) |
#28: RNA chain | Mass: 24223.482 Da / Num. of mol.: 1 / Source method: obtained synthetically Source: (synth.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: GenBank: 176433 |
-Protein , 16 types, 16 molecules CDFKPRSTUZfgJLNX
#2: Protein | Mass: 23162.922 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CKL3 |
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#3: Protein | Mass: 24830.895 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CRK7 |
#4: Protein | Mass: 22912.369 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CRA3 |
#5: Protein | Mass: 11423.085 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CVZ5 |
#7: Protein | Mass: 13220.561 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CKV4 |
#9: Protein | Mass: 14886.174 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CWU3 |
#10: Protein | Mass: 16953.404 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CWT9 |
#11: Protein | Mass: 15747.814 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CXM0 |
#12: Protein | Mass: 12032.146 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CIM1 |
#13: Protein | Mass: 7939.246 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CW78 |
#15: Protein | Mass: 7958.424 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: P69061 |
#16: Protein | Mass: 35612.676 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CNI7 |
#35: Protein | Mass: 20882.344 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CM18 |
#36: Protein | Mass: 17712.734 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CX80 |
#37: Protein | Mass: 16858.678 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CJK0 |
#42: Protein | Mass: 15916.701 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q875M3 |
-40S ribosomal protein ... , 17 types, 17 molecules MQcdeABEGHIOVWYab
#6: Protein | Mass: 12710.457 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CLU4 |
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#8: Protein | Mass: 15656.257 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q875N2 |
#14: Protein | Mass: 6972.152 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: P33285 |
#17: Protein | Mass: 6322.149 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CPG3 |
#18: Protein | Mass: 6608.788 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CUH5 |
#29: Protein | Mass: 24368.662 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CN12 |
#30: Protein | Mass: 26404.725 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CWD0 |
#31: Protein | Mass: 29486.312 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CWJ2 |
#32: Protein | Mass: 25810.977 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CM04 |
#33: Protein | Mass: 21033.570 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CTD6 |
#34: Protein | Mass: 22511.531 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CMG3 |
#38: Protein | Mass: 13458.439 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: P27069 |
#39: Protein | Mass: 9797.949 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CXT6 |
#40: Protein | Mass: 14513.846 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CW21 |
#41: Protein | Mass: 15063.354 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CU44 |
#43: Protein | Mass: 11166.179 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CS01 |
#44: Protein | Mass: 8753.168 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: Q6CNL2 |
-Protein/peptide , 1 types, 1 molecules h
#19: Protein/peptide | Mass: 3354.243 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) References: UniProt: P0CX86 |
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-Eukaryotic translation initiation factor ... , 10 types, 10 molecules imoqkljpsr
#21: Protein | Mass: 12659.412 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Gene: TIF11, YMR260C, YM8156.02C / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: P38912 |
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#22: Protein | Mass: 10299.010 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Gene: SUI1, RFR1, YNL244C, N0905 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: P32911 |
#23: Protein | Mass: 62822.781 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Gene: RPG1, TIF32, YBR079C, YBR0734 / Production host: Saccharomyces cerevisiae (brewer's yeast) / References: UniProt: P38249 |
#24: Protein | Mass: 80545.891 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Gene: NIP1, YMR309C, YM9924.01C, YM9952.11C / Production host: Saccharomyces cerevisiae (brewer's yeast) / References: UniProt: P32497 |
#25: Protein | Mass: 47230.148 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Gene: GCD11, TIF213, YER025W / Production host: Saccharomyces cerevisiae (brewer's yeast) / References: UniProt: P32481 |
#26: Protein | Mass: 16458.090 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Gene: SUI3, TIF212, YPL237W / Production host: Saccharomyces cerevisiae (brewer's yeast) / References: UniProt: P09064 |
#27: Protein | Mass: 30221.979 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Gene: SUI2, TIF211, YJR007W, J1429 / Production host: Saccharomyces cerevisiae (brewer's yeast) / References: UniProt: P20459 |
#45: Protein | Mass: 76738.898 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Gene: PRT1, CDC63, YOR361C / Production host: Saccharomyces cerevisiae (brewer's yeast) / References: UniProt: P06103 |
#46: Protein | Mass: 38229.754 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Gene: TIF34, SCY_4321 / Production host: Saccharomyces cerevisiae (brewer's yeast) / References: UniProt: A6ZMK5, UniProt: P40217*PLUS |
#47: Protein/peptide | Mass: 5576.158 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Gene: TIF35, SCY_1313 / Production host: Saccharomyces cerevisiae (brewer's yeast) / References: UniProt: A6ZZ25 |
-Non-polymers , 4 types, 87 molecules
#48: Chemical | ChemComp-MG / #49: Chemical | ChemComp-ZN / #50: Chemical | ChemComp-GCP / | #51: Chemical | ChemComp-MET / | |
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-Details
Has protein modification | N |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
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Molecular weight | Value: 1.8 MDa / Experimental value: NO | ||||||||||||||||||||||||||||||||||||||||||
Source (natural) |
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Source (recombinant) |
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Buffer solution | pH: 6.5 | ||||||||||||||||||||||||||||||||||||||||||
Buffer component |
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Specimen | Conc.: 0.18 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | ||||||||||||||||||||||||||||||||||||||||||
Vitrification | Instrument: FEI VITROBOT MARK I / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 K |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 59000 X / Nominal defocus max: 3500 nm / Nominal defocus min: 1500 nm / Cs: 2.7 mm / C2 aperture diameter: 70 µm / Alignment procedure: COMA FREE |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Temperature (max): 100 K / Temperature (min): 90 K |
Image recording | Average exposure time: 1.1 sec. / Electron dose: 32 e/Å2 / Detector mode: INTEGRATING / Film or detector model: FEI FALCON II (4k x 4k) / Num. of grids imaged: 3 / Num. of real images: 5500 Details: Images were collected in movie-mode at 32 frames per second |
-Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 1182309 | ||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 5.75 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 12586 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||||||||||
Atomic model building | Protocol: OTHER / Space: RECIPROCAL / Target criteria: FSC |